Caspase-12 Assay Kit, Fluorometric with Ac-ATAD-AFC substrate

Caspase-12 Assay Kit, Fluorometric with Ac-ATAD-AFC substrate

General description

Caspase family of proteases are the central mediators of apoptosis in mammalian cells. The Caspase-12 Fluorometric Assay Kit provides a simple and convenient means for assaying the activity of caspases that recognize the sequence ATAD. The assay is based on detection of cleavage of substrate ATAD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). ATAD-AFC emits blue light (lmax = 400 nm); upon cleavage of the substrate by Caspase-12 or related caspases, free AFC emits a yellow-green fluorescence (λmax = 505 nm), which can be quantified using a fluorometer or a fluorescence microtiter plate reader.

Detection Method:         Fluorescence (Ex/Em 400/505)

Species Reactivity:         Mammalian

Applications:                   Detect early/middle stages of apoptosis; differentiate apoptosis from necrosis

Storage temp.:                -20°C

Features & Benefits:

• Simple one-step procedure that takes only ~1-2 hours
• Fast and convenient
• Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in Caspase-12 activity

Kit Components :

• Cell Lysis Buffer
• 2X Reaction Buffer
• ATAD-AFC Substrate
• DTT (1 M)